self-sufficiency in growth signals limitless replicative potential sustained angiogenesis evading...
TRANSCRIPT
![Page 1: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/1.jpg)
Self-sufficiency in growth signals
Limitless replicative potential
Sustained angiogenesis
Evading apoptosis
Insensitivity to anti-growth signals
Tissue invasion & metastasis
The development of human cancer cells
![Page 2: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/2.jpg)
Cdc6 is essential in the initiation of DNA replication:
A). Assembling pre-replication complex (pre-RC) with ORC, and loading MCMs to replication origins;
B). Licensing DNA replication to ensure one round DNA replication per cell cycle.
![Page 3: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/3.jpg)
The novel function of GLS101/TSUKUSI FL1 is the silencing of human Cdc6, the specific genes for cell proliferation: Specific Cdc6 shRNAs produced from the viral vector Inhibit Cdc6 and blockCancer cell proliferation, but not normal cells.
Cdc6 knockdown
Cdc6 shRNA
Cdc6
TH
TD
6A
i
TH
TP
CHLA255LA-N-2
TH
TP
TH
TD
6A
i
Mo
ck
Mo
ck
TH
TP
TH
TD
6A
i
MCF7
-actin
1 2 3 4 5 6 7 8
![Page 4: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/4.jpg)
G1
S
G2
M
STOP
Recombinant Lentivirus
GLS101
![Page 5: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/5.jpg)
1 2 3 4
Phosph-Rb-C Ser780
Phosph-Rb-C Ser807/811
IgG, light chain
— IP/β-p16INK4a
— — THTD — — — IP/mouse IgG
IgG, light chain
Phosph Rb-C Ser 807/811
1 2 3 4
1:5
1:10
1:25
IP/β-p16INK4a
— — — THTD
Reactivation of p16INK4a CDK inhibitory activity by Cdc6 knockdown
![Page 6: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/6.jpg)
MCF7
MCF7/THTP
MCF7/THTD6Ai
Senescence-associated β-glactosidase (SA-β-Gla) positive staining caused by Cdc6 knockdown
![Page 7: Self-sufficiency in growth signals Limitless replicative potential Sustained angiogenesis Evading apoptosis Insensitivity to anti-growth signals Tissue](https://reader036.vdocuments.us/reader036/viewer/2022070413/5697bfaf1a28abf838c9d308/html5/thumbnails/7.jpg)
Anti-H3K9me3/me2indirect fluorescentstaining
THTD6AiMock
Mock THTD6Ai
Confocal fluorescent microscopy
The formation of senescence associated heterochromatic foci viaCdc6 knockdown